Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

PCR01:32

PCR

228.3K
Overview
228.3K
Real Time RT-PCR02:57

Real Time RT-PCR

61.5K
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
61.5K
Next-generation Sequencing03:00

Next-generation Sequencing

93.2K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
93.2K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Optimisation of Electrokinetic Extraction System: Colourimetric Determination of Copper (II) in Sand Using Polymer Inclusion Membrane.

Electrophoresis·2026
Same author

Redox-mediator enhanced electrochemiluminescence under non-aqueous conditions.

Chemical science·2026
Same author

Rethinking the Excited-State Redox Properties of Iron(III) Complexes for LMCT Photoredox Catalysis.

Journal of the American Chemical Society·2025
Same author

Exploiting a New Strategy to Prepare Water-Soluble Heteroleptic Iridium(III) Complexes to Control Electrochemiluminescence Reaction Pathways in Aqueous Solution.

Chemistry (Weinheim an der Bergstrasse, Germany)·2025
Same author

Maximizing Photon-to-Electron Conversion for Atom Efficient Photoredox Catalysis.

Journal of the American Chemical Society·2024
Same author

Electrochemiluminescence Enhanced by a Non-Emissive Dual Redox Mediator.

Angewandte Chemie (International ed. in English)·2024

Related Experiment Video

Updated: Sep 29, 2025

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
07:35

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems

Published on: June 14, 2021

2.8K

An improved nucleic acid sequence-based amplification method mediated by T4 gene 32 protein.

Yi Heng Nai1, Egan H Doeven1, Rosanne M Guijt1

  • 1Centre for Regional and Rural Futures, Deakin University, Geelong, Victoria, Australia.

Plos One
|March 24, 2022
PubMed
Summary

Incorporating single-stranded binding proteins into Nucleic Acid Sequence-Based Amplification (NASBA) simplifies workflows. T4 gene gp32 protein enhanced NASBA efficiency for HIV-1 RNA detection, showing potential for point-of-care applications.

More Related Videos

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
05:41

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications

Published on: July 10, 2020

2.1K
Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
15:49

Reverse Genetics to Engineer Positive-Sense RNA Virus Variants

Published on: June 9, 2022

1.5K

Related Experiment Videos

Last Updated: Sep 29, 2025

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
07:35

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems

Published on: June 14, 2021

2.8K
2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
05:41

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications

Published on: July 10, 2020

2.1K
Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
15:49

Reverse Genetics to Engineer Positive-Sense RNA Virus Variants

Published on: June 9, 2022

1.5K

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostics

Background:

  • Nucleic Acid Sequence-Based Amplification (NASBA) is crucial for isothermal amplification but often requires a complex thermal denaturation step.
  • Simplifying NASBA workflows is essential for its adoption in point-of-care (POC) testing.
  • Single-stranded binding proteins (SSBs) have enhanced other amplification methods, but their effect on NASBA is underexplored.

Purpose of the Study:

  • To investigate the efficacy of incorporating three different SSBs (RecA, ET SSB, gp32) into a one-step NASBA protocol.
  • To evaluate the impact of SSBs on amplification efficiency and workflow simplification for NASBA.
  • To assess the potential of SSB-modified NASBA for sensitive and rapid detection of viral RNA, such as HIV-1.

Main Methods:

  • Three SSBs (RecA, ET SSB, gp32) were integrated into a one-step, single-pot NASBA assay performed at 41 °C.
  • The modified NASBA protocol was tested for amplification of synthetic HIV-1 RNA.
  • Amplification efficiency and time-to-positivity (ttp) were compared against conventional two-step NASBA and one-step NASBA without SSBs.

Main Results:

  • All tested SSBs significantly improved NASBA amplification compared to the standard two-step process.
  • T4 gene gp32 protein (gp32) demonstrated improved amplification without non-specific amplification.
  • Gp32 reduced the time-to-positivity for synthetic HIV-1 RNA detection by an average of 13.6% in one-step NASBA and 6.7% in conventional NASBA.

Conclusions:

  • Single-stranded binding proteins, particularly gp32, can enhance NASBA performance and simplify its workflow.
  • The use of gp32 in NASBA shows promise for improving the speed and efficiency of RNA detection.
  • This SSB-mediated enhancement supports the development of simplified NASBA protocols for point-of-care diagnostic applications.